1. Field of the Invention
The present invention relates to a method for determining a genotoxicity using non-fluorescent fluorescence proteins.
2. Description of the Related Art
The introduction of chemicals into the environment represents a serious risk to environmental and human health. Therefore, current legislation in European and other industrial countries requires appropriate data on risk assessment for the registration of chemicals, pesticides, biocides and pharmaceuticals (Commission of the European Communities 1967, 1991, 1992, 1993a, b, 1994; CVMP/VICH 2000; EMEA/CHMP 2006; VICH 2004). Although various genotoxicity assays have been developed to assess toxicity of these chemicals, by far the most frequently used is the Ames assay (Ames, Durston, et al., 1973; Maron and Ames, 1983). This assay is based on the detection of reversal of point mutations in the his gene of a defective mutant of Salmonella typhimurium by quantifying the growth in cultures without histidine. The Ames assay has the critical disadvantage that Salmonella does not possess the cellular machinery of vertebrates which is necessary to convert procarcinogens into reactive metabolites. Thus, several transgenic animal systems including MutaMouse (Gossen, de Leeuw, et al., 1989), BigBlue mouse (Kohler, Provost, et al., 1991), gpt-Δ transgenic mouse systems (Nohmi, Katoh, et al., 1996), and rpsL transgenic zebrafish (Amanuma, Takeda, et al., 2000) have been developed. In all cases, however, transgenic reporter genes, all with prokaryotic origin, must be introduced and assessed in appropriate test bacteria, E. coli, after extraction of DNA from animal organs.
Throughout this application, various publications and patents are referred and citations are provided in parentheses. The disclosures of these publications and patents in their entities are hereby incorporated by references into this application in order to fully describe this invention and the state of the art to which this invention pertains.